miR-181a抑制舌鳞癌细胞上皮-间充质转化及化疗耐药机制的研究

doi:10.19438/j.cjoms.2018.04.001

中国口腔颌面外科杂志 ›› 2018, Vol. 16 ›› Issue (4): 289-295.doi: 10.19438/j.cjoms.2018.04.001

miR-181a抑制舌鳞癌细胞上皮-间充质转化及化疗耐药机制的研究

刘墨¹, 李劲松¹, 阮毅¹, 黄洪章²

1.中山大学孙逸仙纪念医院口腔科,广东广州 510120;
2.中山大学光华口腔医学院口腔颌面外科,广东广州 510055

收稿日期:2017-09-08 修回日期:2018-01-17 出版日期:2018-07-20 发布日期:2018-08-09
通讯作者: 黄洪章,E-mail:huanghongzhang@tom.com
作者简介:刘墨(1978-),女,博士,副主任医师,E-mail：mmliumo@163.com
基金资助:
国家自然科学基金 (81502350）; 广东省基础与应用基础研究专项（省自然基金）自由申请项目（2016A030313352）

Study on miR-181a inhibiting epithelial-mesenchymal transition and drug-resistance by targeting Twist1 in tongue squamous cell carcinoma

LIU Mo¹, LI Jin-song¹, RUAN Yi¹, HUANG Hong-zhang²

1.Department of Stomatology, Sun Yat-sen Memorial Hospital, Sun Yat-sen University. Guangzhou 510120;
2.Department of Oral and Maxillofacial Surgery, Guanghua School and Research Institute of Stomatology, Sun Yat-sen University. Guangzhou 510055, Guangdong Province, China

Received:2017-09-08 Revised:2018-01-17 Online:2018-07-20 Published:2018-08-09

摘要/Abstract

摘要： 目的:探讨miR-181a调控上皮-间充质转化(EMT)促进舌鳞状细胞癌细胞顺铂耐药的机制。方法:分别在CAL27/CDDP、CAL27中转染miR-181a mimics、miR-181a LNA,免疫印迹及免疫荧光检测E-cadherin、Vimentin的表达;免疫印迹检测Twist1、Slug的表达;Transwell及划痕实验检测细胞的侵袭、迁移能力;MTT法检测细胞对顺铂的半抑制率。利用生物信息学方法:预测Twist1可能为miR-181a的靶基因。构建包含Twist1结合序列片段的荧光素酶报告基因质粒,进行双荧光素酶报告基因实验。采用SPSS17.0软件包对结果进行单因素方差分析。结果:转染miR-181a mimics可逆转CAL27/CDDP的EMT表型,E-cadherin表达增强,Vimentin、Twist1及Slug表达降低,细胞的侵袭、迁移能力显著降低,IC₅₀下降47.57%±6.23%（P<0.05）。转染miR-181a LNA可诱导CAL27细胞发生EMT,E-cadherin表达降低,Vimentin、Twist1及Slug表达增强,细胞侵袭、迁移能力显著增强,IC₅₀升高55.61%±7.20%（P<0.05）。双荧光素酶报告基因实验证实,Twist1是miR-181a的靶基因。结论:miR-181a靶向调控Twist1,从而抑制舌鳞癌细胞上皮-间充质转化与顺铂耐药。

关键词: 舌鳞状细胞癌, 上皮-间充质转化, microRNA, Twist1, 耐药性

Abstract: PURPOSE:The aim of this study was to investigate the mechanism of miR-181a regulating epithelial-mesenchymal transition (EMT) and enhancing the capacity of cisplatin-resistance of tongue squamous cell carcinoma cells. METHODS: TSCC CDDP resistant cell line CAL27/CDDP and its parent cell line CAL27 were transfected with miR-181a mimics and miR-181a LNA respectively. Western blot and immunofluorescence staining were used to detect the expression of E-cadherin and Vimentin; Western blot was used to detect the expression of Twist1 and Slug; Transwell assay and scratch test were used to detect the invasion and migration capabilities; MTT was used to analyze the half maximal inhibitory concentration (IC₅₀) values. Bioinformatics analysis was performed and found that Twist1 was directly targeted by miR-181a. Luciferase reporter gene plasmids were constructed and designed obligos including miR-181a targeting site. Dual luciferase reporter gene array was performed. SPSS17.0 software package was used to analyze the results. RESULTS: miR-181a mimics transfection reversed the EMT phenotype of CAL27/CDDP. The expression of E-cadherin was up-regulated and the expression of Vimentin, Twist1 and Slug was down-regulated. The motility was significantly decreased. IC₅₀ of CAL27/CDDP decreased by 47.57%±6.23% (P<0.05). miR-181a LNA transfection induced EMT in CAL27. The expression of E-cadherin was down-regulated and the expression of Vimentin, Twist1 and Slug was up-regulated. The motility was significantly increased. IC₅₀ of CAL27 increased by 55.61%±7.20%(P<0.05).CONCLUSIONS: MiR-181a inhibits EMT and CDDP drug-resistance by directly targeting Twist1 in TSCC.

Key words: Tongue squamous cell carcinoma, Epithelial-mesenchymal transition, microRNA, Twist1, Drug resistance

中图分类号:

R739.8

刘墨, 李劲松, 阮毅, 黄洪章. miR-181a抑制舌鳞癌细胞上皮-间充质转化及化疗耐药机制的研究[J]. 中国口腔颌面外科杂志, 2018, 16(4): 289-295.

LIU Mo, LI Jin-song, RUAN Yi, HUANG Hong-zhang. Study on miR-181a inhibiting epithelial-mesenchymal transition and drug-resistance by targeting Twist1 in tongue squamous cell carcinoma[J]. China J Oral Maxillofac Surg, 2018, 16(4): 289-295.

参考文献

[1] Li S, Zhang X, Zhang R, et al.Hippo pathway contributes to cisplatin resistant-induced EMT in nasopharyngeal carcinoma cells[J]. Cell Cycle, 2017, 16(17): 1601-1610.
[2] Maciaczyk D, Picard D, Zhao L, et al.CBF1 is clinically prognostic and serves as a target to block cellular invasion and chemoresistance of EMT-like glioblastoma cells[J]. Br J Cancer, 2017, 117(1): 102-112.
[3] Rice AJ, Cortes E, Lachowski D, et al.Matrix stiffness induces epithelial-mesenchymal transition and promotes chemoresistance in pancreatic cancer cells[J]. Oncogenesis, 2017, 6(7): e352.
[4] Pang MF, Georgoudaki AM, Lambut L, et al.TGF-β1-induced EMT promotes targeted migration of breast cancer cells through the lymphatic system by the activation of CCR7/CCL21-mediated chemotaxis[J]. Oncogene, 2016, 35(6): 748-760.
[5] 刘墨, 张斌, 王安训, 等. 人舌鳞状细胞癌顺铂耐药细胞发生上皮-间质转化的研究[J].中华口腔医学研究杂志:电子版, 2014, 8(5): 357-363.
[6] 刘墨, 张斌, 王成, 等. 转化生长因子β1诱导人舌鳞状细胞癌细胞上皮-间质转化及其顺铂耐药性研究[J]. 中华口腔医学研究杂志:电子版, 2012, 6(3): 223-231.
[7] 刘墨, 阮毅, 李劲松, 等. Twist1调控上皮-间质转化促进舌鳞癌细胞顺铂耐药机制的研究[J].中国口腔颌面外科杂志, 2018, 16(1): 1-5.
[8] 周晓健, 张志愿, 张萍, 等. 舌鳞状细胞癌顺铂耐药细胞系细胞周期蛋白D1RNA干扰的体外研究[J].中华口腔医学杂志, 2006, 41(11): 646-649.
[9] 何春梅, 段朝军, 泰迪兰, 等. 维拉帕米对人舌癌耐药细胞株Tca8113/BLM的逆转作用研究[J]. 中国现代医学杂志, 2008, 18(4): 417-419.
[10] 冯戈, 王大章, 陈槐卿, 等. 基因芯片对荷瘤裸鼠Tca8113/卡铂耐药基因表达谱的分析[J]. 实用口腔医学杂志, 2007, 23(5): 688-691.
[11] Sun L, Yao Y, Liu B, et al.MiR-200b and miR-15b regulate chemotherapy-induced epithelial-mesenchymal transition in human tongue cancer cells by targeting BMI1[J]. Oncogene, 2012, 31(4): 432-445.
[12] Dai X, Fang M, Li S, et al.miR-21 is involved in transforming growth factor β1-induced chemoresistance and invasion by targeting PTEN in breast cancer[J]. Oncol Lett, 2017, 14(6): 6929-6936.
[13] Yang RM, Zhan M, Xu SW, et al.miR-3656 expression enhances the chemosensitivity of pancreatic cancer to gemcitabine through modulation of the RHOF/EMT axis[J]. Cell Death Dis, 2017, 8(10): e3129.

[1]	鲁煜, 刘清海, 梁海胜, 邓刚. 3D-ASL对舌肿物的诊断价值及与病理学特征的相关性分析[J]. 中国口腔颌面外科杂志, 2024, 22(1): 42-48.
[2]	王博, 沈月洪, 杨宏宇. 鼻咽癌放化疗后继发舌鳞癌、口咽癌及牙龈癌1例并文献复习[J]. 中国口腔颌面外科杂志, 2023, 21(3): 301-305.
[3]	水文军, 高金星, 任蕾. 基质金属蛋白酶9抑制剂对人舌鳞癌SCC-25细胞侵袭、迁移能力的影响及机制探讨[J]. 中国口腔颌面外科杂志, 2021, 19(5): 424-428.
[4]	陈正岗, 王奇民, 童磊, 王云英, 徐晓娜, 王莹, 韩红钰, 盛善桂, 王少如. 沉默Icmt基因对舌鳞癌CAL-27和SCC-4细胞增殖、凋亡及细胞周期的影响[J]. 中国口腔颌面外科杂志, 2021, 19(2): 97-104.
[5]	孔宪琛, 姜晓东, 芦昕, 禹雯怡, 薛娇, 张鹏, 赵璐, 袁荣涛. 核糖体蛋白L29在舌鳞状细胞癌中的表达及意义[J]. 中国口腔颌面外科杂志, 2021, 19(2): 110-115.
[6]	王子昱, 丁宇洁, 陈冰, 沈昊翀, 孟箭, 仝伟凤. α桐酸对舌鳞状细胞癌CAL-27细胞的体外抑制作用[J]. 中国口腔颌面外科杂志, 2020, 18(6): 490-494.
[7]	周光明, 梁衍灿, 黄子贤, 张彬. 长链非编码RNA FOXD2-AS1对舌鳞癌细胞迁移、侵袭的影响[J]. 中国口腔颌面外科杂志, 2020, 18(6): 495-500.
[8]	徐桥石, 杨溪, 马海龙, 常涵月, 张陈平. 白细胞介素8在口腔鳞癌上皮-间充质转化中的研究进展[J]. 中国口腔颌面外科杂志, 2020, 18(6): 563-566.
[9]	陈永菊, 陈伟良, 周斌, 钟江龙, 汪延, 陈睿. 面-颏下动脉岛状皮瓣修复年轻与老年舌癌患者的效果比较[J]. 中国口腔颌面外科杂志, 2020, 18(2): 117-121.
[10]	刘国新, 王伟伟, 麦潋曦, 焦九阳, 谢舒乐, 林钊宇. 顺铂诱导舌鳞癌细胞发生上皮-间充质转化及获得肿瘤干细胞样特性的实验研究[J]. 中国口腔颌面外科杂志, 2020, 18(1): 10-16.
[11]	林志立, 梁建平, 周斌. lncRNA-HOTTIP在低氧诱导舌鳞癌细胞上皮-间质转化过程中的作用探讨[J]. 中国口腔颌面外科杂志, 2019, 17(1): 26-31.
[12]	罗丹, 孙巧珍, 刘少华, 孙俊伟, 王升志. 热疗联合Id-1基因沉默对舌鳞癌CAL-27细胞增殖及侵袭的影响[J]. 中国口腔颌面外科杂志, 2018, 16(6): 488-492.
[13]	马利, 石冰, 张东升, 郑谦. 细胞凋亡及上皮-间充质转化在腭胚突融合过程中的作用探讨[J]. 中国口腔颌面外科杂志, 2018, 16(6): 493-497.
[14]	卢陈佩, 李洪亮, 欧阳宁鹃, 林雨恒, 司家文, 沈国芳. MicroRNA-103-3p对小鼠前成骨细胞MC3T3-E1成骨分化早期的影响[J]. 中国口腔颌面外科杂志, 2018, 16(4): 302-309.
[15]	王超, 黎婷, 严斐, 蔡文燕, 蒋星宇, 孙晋虎. 辛伐他汀联合干扰miR-21对唾液腺腺样囊性癌细胞迁移和侵袭的影响[J]. 中国口腔颌面外科杂志, 2018, 16(2): 132-137.

miR-181a抑制舌鳞癌细胞上皮-间充质转化及化疗耐药机制的研究

Study on miR-181a inhibiting epithelial-mesenchymal transition and drug-resistance by targeting Twist1 in tongue squamous cell carcinoma

可视化

摘要/Abstract

引用本文

使用本文

参考文献

相关文章 15

编辑推荐

Metrics

本文评价